The intent of this project is to develop improved methods for the cryopreservation of human cancer cell lines and tumor biopsies. While there has been a current emphasis on the importance of improving biobanking of tissues (GEN, 2/1/05), there has been little research dedicated to improving the archival storage of cancer samples. Cell Preservation Services Inc. (CPSI) develops hypothermic storage (HypoThermosol, "HTS") and cryopreservation (CryoStor) solutions marketed by CPSI's partner, BioLife Solutions. These solutions are used as the shipping/storage solutions in Regenerative Medicine applications such as cellular cardiomyoplasty. The fully defined, serum-free CryoStor platform is designed for cell and tissue storage in liquid nitrogen, requires reduced DMSO levels, and improves cryopreservation efficacy up to 50% compared to traditional protocols. Yet, even the CryoStor series can only protect the viability of cancer cells to a maximum of 70%. Thus, this "cryopreservation cap" must be attacked so as to achieve optimal preservation. CPSI is able to develop improved HTS and CryoStor solutions given its ability to investigate and modulate the cell death cascades that can be initiated due to extended storage. The purpose of this Phase I Project is to develop a new platform of solutions called CryoStor-CANCER - a series of solutions expressly designed for the improved cryopreservation of human cancer cells and tumor biopsies. CPSI will (1) use cDNA microarrays to determine if the analysis of stress pathways activated by cancer cells undergoing cryopreservation can lead to improved preservation solutions for cancer cells and tumor biopsies; (2) determine if the same analysis of stress pathways might lead to "rescue solutions" that can reverse the adverse effects of sub-optimal cryopreservation and; (3) determine if sub-optimal preservation results in a long term change to the expression of any of the 400+ genes that are most often studied in cancer biology. Phase 2 studies will test the prototype CryoStor solutions on a wider variety of cancer cell and tumor types, and determine if cryopreservation and the proposed "rescue solution" preserve or rescue the oncological fingerprint of cancer cell lines and tumor tissues. This work will be important to agencies that are currently developing improved methods for biobanking of cryopreserved cancer specimens. As a result, archival storage of cancer tissues will be improved and poorly preserved specimens can be rescued. [unreadable] [unreadable] [unreadable]